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目的:探讨自拟方龙蛭汤对气虚血瘀证脑梗死大鼠急性期神经功能缺损评分、血管新生及促血管生成因子HIF-1α(hypoxia-inducible factor-1α,HIF-1α)、促血管生成素-2(angiopoietin-2,Ang-2)水平的影响。方法:60只SD大鼠随机分为模型组、对照组和治疗组,每组各20只。3组均采用力竭游泳14天+改良线栓法建立气虚血瘀证脑梗死模型,造模成功后分别予生理盐水、补阳还五汤和龙蛭汤灌胃并比较给药前、第7、14天神经功能缺损评分(zea-longa评分)。在第7、14天两时间点取大鼠缺血脑组织采用免疫组织化学法和Western blot法检测CD34、HIF-1α、Ang-2蛋白表达水平,并测定微血管计数(microvessel count,MVC)。结果:(1)给药前3组大鼠zea-longa评分无明显差异(均P>0.05),第7、14天对照组和治疗组均明显低于模型组(均P<0.05)。(2)免疫组化结果显示:第7、14天治疗组MVC、HIF-1α、Ang-2均明显高于模型组,且高于对照组(均P<0.05)。(3)Western blot结果显示:第7、14天,治疗组CD34、Ang-2、HIF-1α均明显高于模型组,且高于对照组(均P<0.05)。结论:龙蛭汤能促进气虚血瘀证脑梗死大鼠急性期神经功能恢复和缺血脑组织血管新生,其机制可能与上调促血管生成因子HIF-1α、Ang-2相关。
Objective: To investigate the effects of Fanglongzhihu decoction on acute neurological deficit score, angiogenesis and hypoxia-inducible factor-1alpha (HIF-1alpha), pro-angiogenesis Angiopoietin-2 (Ang-2) levels. Methods: Sixty SD rats were randomly divided into model group, control group and treatment group, 20 rats in each group. The rats in each group were treated with exhausted swimming for 14 days and modified suture method to establish cerebral infarction model of Qi-deficiency and Blood-stasis syndrome. After the model was established successfully, the rats were given normal saline, Bu Yang Huan Wu Decoction and Longzhi Decoction respectively, Neurological deficit scores at 7 and 14 days (zea-longa score). At the 7th and 14th days, the ischemic brain tissue was taken at two time points. The expressions of CD34, HIF-1α and Ang-2 were detected by immunohistochemistry and Western blot, and the microvessel count (MVC) was measured. Results: (1) There was no significant difference in the zea-longa score between the three groups before administration (all P> 0.05). On the 7th and 14th days, the zea-longa scores in the control and treatment groups were significantly lower than those in the model group (all P <0.05). (2) The results of immunohistochemistry showed that MVC, HIF-1α and Ang-2 in the treatment group were significantly higher than those in the model group on the 7th and 14th days (all P <0.05). (3) Western blot results showed that CD34, Ang-2 and HIF-1α in the treatment group were significantly higher than those in the model group on the 7th and 14th days (all P <0.05). Conclusion: Longzhi Decoction can promote neurological recovery and ischemic brain angiogenesis in acute stage of cerebral infarction with qi deficiency and blood stasis syndrome in rats. The mechanism may be related to the up-regulation of angiogenic factors HIF-1α and Ang-2.